Enkephalin, a natural opiate receptor agonist in the brain, has been identified [see Hughes et al., Nature, 256 577 (1976)] as a mixture of two pentapeptides: H-Tyr-Gly-Gly-Phe-Met-OH (methionine-enkephalin) and H-Tyr-Gly-Gly-Phe-Leu-OH (leucine-enkephalin). Both peptides mimic the ability of morphine to block electrically evoked contractions of mouse vas deferens and guinea pig ileum, and both inhibit the sterospecific receptor binding of the opiate antagonist 3H-naloxone in brain homogenates.
It has been proposed that enkephalin receptors may be sites at which morphine-like drugs exert their analgesic activities, and that enkephalin may be the modulator or transmittor in brain systems for pain suppression or analgesia. It has been reported that methionine-enkephalin and leucine-enkephalin, when administered by injection in the brain ventricle in rats, induce a profound analgesia that is fully reversible by naloxone. [See Beluzzi et al., Nature, 260, 625 (1976)]. The naturally occurring enkephalins are inactive when administered peripherally, presumably because they are rapidly destroyed by blood enzymes and/or are poorly transported across the blood-brain barrier.
The amino acid sequence of methionine-enkephalin is identical to that of the N-terminal portion of the C-fragment (.beta.-endorphin or .alpha.-LPH[61-91]) of the peptide .beta.-lipotropin, which is found in large concentrations in the pituitary and in much lower concentrations in the brain. Other naturally-occurring fragments of .beta.-lipotropin are known, for example: .alpha.-endorphin (.beta.-LPH[61-76]) and .gamma.-endorphin (.beta.-LPH[61-77]). Both .beta.lipotropin and the endorphins show morphine-like properties in various test systems, and it has been suggested that methionine-enkephalin is a breakdown product of the large opiate-like peptides. Enkephalin, its relationship to .beta.-lipotropin and the endorphins, and the pharmacological properties thereof, are reviewed in an article by Iversen et al., Nature, 262, 738 (1976). Recent developments are also described in detail in the "Proceedings of the International Narcotics Research Club Meeting, Abderdeen, U.K., July 19-22, 1976," published in OPIATES AND ENDOGENOUS OPIOID PEPTIDES, North Holland Publishing Company, Amsterdam, 1976.
Various structural variations of methionine-enkephalin and leucine-enkephalin are described in the literature. For example, the pentapeptide H-Tyr-Gly-Gly-Phe-Thr-OH, wherein the fifth amino acid residue (methionine or leucine) is replaced by threonine, is described by Chang et al., Life Sciences, 18, 1473(1976). Similarly, a long acting synthetic pentapeptide, Tyr-D-Ala-Gly-Phe-Met-amide is described in Pert et al., Science, 194, 330 (1976); which compound, like the natural enkephalins, is reportedly inactive when administered peripherally. Baxter et al., British Journal of Pharmacology, March 2, 1977, pages 455P-456P and 523P report activity in the compound Tyr-D-Ala-Gly-Phe-D-Leu when administered intracerebroventricularly. Coy et al. B.B.R.C. 73 632 (1976) disclose that D-Met.sup.5 enkephalin has one tenth the activity of Met enkephalin. Bajusz et al. FEBS Letters 76, 91 (1977) by replacing Gly.sup.2 with D-Met and the Met.sup.5 by Pro-NH.sub.2 obtained a very potent antinociceptive pentapeptide Tyr-D-Met-Gly-Phe-Pro-NH.sub.2 which was 5.5 times more potent than morphine by intravenous administration.
Romer et al., Nature, 268 547 (1977) showed that the substituted tetrapeptide amide ##STR2## possesses potent peripheral analgesic activity and some analgesic activity when given orally at high doses (200-300 mg/kg).
Morgan et al., "Peptides", Proc. Fifth Amer. Pept. Symp. ed. Goodman and Meienhofer, p. 111 (1977) reported in vitro and in vivo biological activities of several enkephalin analogs among which was N(Me)Tyr-Gly-Gly-Phe-Met-NH-Propyl. Ling et al., ibid., p. 96 (1977) reported in vitro activities of several analogs of enkephalin with D-amino acids in position 5.
Dutta et al., Life Sciences 21, 559 (1977) and Dutta et al., Acta. Pharm. Science 14, 14 (1977) described several analogs with D-Ser, D-Met, D-Ala, D-Thr, D-Lys(Boc), D-Phe, D-Leu, D-Asp and D-Ser(t-Bu), at position 2 and various substitutions with L-amino acids or amines at position 5.
U.S. Pat. No. 4,148,786 discloses: ##STR3##
Belluzzi et al., Life Sciences, 23, 99 (1978) described analogs with D-Ala at position 2 and D-Leu or D-Met at position 5.